1.Influlance of different drying methods on quality of Schisandrae Chinensis Fructus.
Kai-Long AN ; De-Kun LI ; Da-Zheng ZHOU ; Zheng-Liang YE ; Qiao-Sheng GUO
China Journal of Chinese Materia Medica 2014;39(15):2900-2906
OBJECTIVETo study the influence of different drying methods on the quality of Schisandrae Chinensis Fructus and thus provide useful reference for its proper drying methods.
METHODSchisandrae Chinensis Fructus was processed by eight drying methods including vacuum freeze drying, natural drying in the shade, drying in the sun, oven drying and vacuum drying under different temperature. The contents of the functional ingredients includes chisandrin, gomisin D, gomisin J, schisandrol B, angeloylgomisin H, angeloylgomisin Q, gomisin G, schisantherin A, deoxyschisandrin, schisandrin B, schisandrin C, 5-HMF, total aids and total sugars. The main components change after drying were analyzed by HPLC, ultraviolet spectrophotometry and potentiometric titration. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Schisandrae Chinensis Fructus.
RESULTAll these results are in accordance with the requirements of Chinese Pharmacopoeia published in 2010, the contents of schisandrin and total eleven lignans were the highest using vacuum drying, and 5-HMF were the lower, oven drying made little difference but with lower schisandrin and higher 5-HMF as the heat increased.
CONCLUSIONDifferent drying methods have significant influence on the quality of Schisandrae Chinensis Fructus. Oven drying under 5°C should be adopted to substitute drying in the sun according to the China Pharmacopoeia published in 2010 for Schisandrae Chinensis Fructus by comprehensive analysis of the cost, content and practicality.
Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Schisandra ; chemistry ; Temperature
2.One stage correction of sunken eyes combined with ptosis.
Cui-yun LIU ; Da ZHOU ; Kai LIU
Chinese Journal of Plastic Surgery 2012;28(6):424-427
OBJECTIVETo investigate the technique and therapeutic effect for correction of sunken eyes combined with ptosis.
METHODSIn order to adjust the levator muscle tension and the relationship between levator aponeurosis and tarsus plate, multiple individualized treatment was selected, including levator aponeurosis restoration, levator aponeurosis plication, or shorten, or combination. Then the orbital fat was transferred to the depressed area, or autologous fat particles were collected and injected into the depressed area within the orbital fat fascia. After the orbital septum fascia was restored, the incision was closed primarily.
RESULTS15 cases (30 eyes) were treated. 11 cases were followed up for 6-40 months (average, 9.5 months) with satisfactory cosmetic and functional result. No recurrence of ptosis happened.
CONCLUSIONSOne-stage correction of sunken eyes combined with ptosis can be achieved with autologous fat injection or orbital fat transposition. Good cosmetic and functional result can be achieved.
Adipose Tissue ; transplantation ; Aged ; Blepharoplasty ; methods ; Blepharoptosis ; surgery ; Eyelids ; Fasciotomy ; Humans ; Oculomotor Muscles ; surgery ; Orbit
3.Utilization of a Microsateilite Marker to Identify Rice Blast Resistance Gene in Two Segregating Populations
Shi-Gui LI ; Yu-Ping WANG ; Han-Yun LI ; Kai-Da ZHOU ; Li-Huang ZHU
Chinese Journal of Biotechnology 2000;16(3):324-327
The microsatellite (SSLP) marker RM262,which is tightly linked to the blast resistance gene Pi-d(t) in rice, provides means to conduct marker-aided selection in a rice breeding program. The objective of this study was to investigate the ability of this marker to select the blast resistance gene, Pi-d(t), in two crosses for rice blast resistance breeding. The products with amplified the microsatellite primers were polymorphic between the three varietes examined. To examine the power of the identified microsatellite marker in predicting the Pi-d(t) locus, we determined the genotypes of the two F2 populations at the Pi-d(t) locus by performing progeny testing for the disease respones. The results indicated an accuracy of more than 98 % in identifying the resistant plants in both populations. Therefore the microsatellite marker can be utilized in marker-assisted selection and breeding for new varieties with blast resistance.
4.Cloning and prokaryotic expression of major ampullate spidroin gene of spider.
Hong-Chun PAN ; Da-Xiang SONG ; Kai-Ya ZHOU ; Guo-Ping ZHU
Chinese Journal of Biotechnology 2007;23(3):446-451
RT-PCR was conducted with one degenerate primer designed according to repetitive regions' amino acid sequence of major ampullate spidroin (MaSp) in spiders and adaptor primer in the SMART cDNA Library Construction Kit. By cloning and sequencing of amplified products, one cDNA clone (GenBank Accession No. AY365017) of Argiope amoena MaSp gene was obtained. The deduced amino acid sequence can be distinctly divided into two regions: (1) Repetitive region that consists of an alternating alanine-rich and glycine-rich domain in which many prolines are present; and (2) C-terminal non-repetitive region. The region coding for 272 amino acids of MaSp gene was subcloned into prokaryotic expression vector pET28b(+) and an about 26kD recombinant protein was expressed at high levels in Escherichia coli BL21 (DE3) after induction of IPTG. After being purified with metal-affinity chromatography on Ni(2+) -IDA-Sepharose columns as well as gel filtration chromatography, the recombinant protein was confirmed to be predicted MaSp by means of amino acid composition analysis and N-terminal amino acid sequence analysis. The solubility behavior of recombinant MaSp with C-terminal non-repetitive region in the present study is similar to that of recombinant dragline silk proteins without C-terminal non-repetitive region expressed by bacteria and yeast in the other studies. The result shows that absence or presence of C-terminal non-repetitive region is not a crucial factor affecting the solubility of the recombinant MaSp.
Amino Acid Sequence
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Animals
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Base Sequence
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Chromatography, Affinity
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Cloning, Molecular
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DNA, Complementary
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chemistry
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genetics
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli
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genetics
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Fibroins
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genetics
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metabolism
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Gene Expression
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Molecular Sequence Data
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Molecular Weight
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Plasmids
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genetics
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Recombinant Proteins
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chemistry
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isolation & purification
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metabolism
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Sequence Analysis, DNA
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Sequence Analysis, Protein
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Sequence Homology, Amino Acid
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Spiders
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genetics
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metabolism
5.Lithium chloride combined with human umbilical cord blood mesenchymal stem cell transplantation for treatment of spinal cord injury in rats.
Xu-yong DENG ; Rong-ping ZHOU ; Kai-wu LU ; Da-di JIN
Journal of Southern Medical University 2010;30(11):2436-2439
OBJECTIVETo observe the effects of lithium chloride combined with human umbilical cord blood mesenchymal stem cell (hUCB-SCs) transplantation in the treatment of spinal cord injury in rats.
METHODSEighty female SD rats with complete T9 spinal cord transaction were randomized into 4 groups (n=20), namely the control group (group A), lithium chloride group (group B), hUCB-SCs group (group C) and hUCB-SCs(+) lithium chloride group (group D). On days 1 and 3 and the last days of the following weeks postoperatively, the motor function of the hindlimb of the rats were evaluated according to the BBB scores. At 8 weeks, all the rats were sacrificed and the spinal cords were taken for morphological observation. The spinal cord tissues at the injury site were observed with Brdu nuclear labeling to identify the survival and migration of the transplanted SCs. The regeneration and distribution of the spinal nerve fibers were observed with fluorescent-gold (FG) spinal cord retrograde tracing.
RESULTSBrdu labeling showed that the transplanted hUCB-SCs survived and migrated in the spinal cord 8 weeks postoperatively in groups C and D. FG retrograde tracing identified a small amount of pyramidal cells that migrated across the injury site in groups C and D. The BBB scores of the hindlimb motor function 8 weeks postoperatively were 4.11∓0.14, 4.50∓0.15, 8.31∓0.11 and 11.15∓0.18 in groups A, B, C and D, respectively.
CONCLUSIONLithium chloride can promote the survival and differentiation of hUCB-SCs into neural cells at the injury site. Lithium chloride combined with hUCB-SCs transplantation may accelerate functional recovery of the hindlimbs in rats with complete transection of the spinal cord.
Animals ; Cord Blood Stem Cell Transplantation ; Female ; Humans ; Lithium Chloride ; therapeutic use ; Rats ; Spinal Cord Injuries ; therapy
6.Anther culture generated stem borer-resistance DH lines of Minghui 81(Oryza sativa L. subsp. indica) expressing modified cry1Ac gene.
Qian-Chun ZENG ; Qian WU ; De-Jiang FENG ; Kai-Da ZHOU ; Xiang LIU ; Zhen ZHU
Chinese Journal of Biotechnology 2002;18(4):442-446
2600 Anthers from T0 modified cry1 Ac-transgenic rice lines of Minghui 81, an elite restoring line of commercial CMS indica hybrid rice, were cultured on SK3 media. 83 green plantlets were recovered, 43 double haploid (DH) and 40 haploid among them. Results of PCR analyzes indicated that 55 plants of 83 were harbored the cry1Ac gene, and the ratio of cry1Ac-positive against cry1Ac-negative was 2:1 (55/28). 36 putative transgenic DH plants were further confirmed by Southern blot. ELISA detection showed that Cry1Ac level in different transgenic rice plants of the same cry1Ac-DH clone was almost equal and the highest one amount to 0.25% of the total soluble protein. Pest insect-resistant bioassay at field trials demonstrated that some of the homozygous cry1Ac-transgenic rice plants not only showed high-level resistance against striped stem borer (Chilo suppressalis) but also retained elite agronomy characters. These results demonstrated that rice anther culture has a great value in rice molecular breeding.
Animals
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Bacillus thuringiensis
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genetics
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Bacterial Proteins
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genetics
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metabolism
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Bacterial Toxins
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Blotting, Southern
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Culture Techniques
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DNA, Plant
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genetics
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Endotoxins
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genetics
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metabolism
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Enzyme-Linked Immunosorbent Assay
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Gene Expression
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Hemolysin Proteins
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Immunity, Innate
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genetics
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Moths
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growth & development
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Oryza
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genetics
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growth & development
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parasitology
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Plant Structures
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genetics
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growth & development
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parasitology
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Plants, Genetically Modified
7.Multi-plane hyaluronic acid injection for rhinoplasty.
Ren BU ; Da ZHOU ; Ke XUE ; Cheng-An JIANG ; Bin GU ; Kai LIU
Chinese Journal of Plastic Surgery 2012;28(5):337-340
OBJECTIVETo investigate the effect of multi-plane Hyaluronic acid (HA) injection for rhinoplasty.
METHODSThe HA was injected below or above the periosteum at the nasal bone, above the perichondrium at the cartilage portion of nose, and between the great alar cartilage at the nasal tip. The HA volume was 1-1.5 ml, according to the nose form and aesthetic assessment. Over-injection was not permitted. Touch-up injection could be performed one week after the first injection if need.
RESULTSFrom Jan. 2010 to Jan. 2012, 60 cases underwent rhinoplasty with HA injection. The patients were followed up for 10-13 months with satisfactory result. The effect lasted about 9 months with the longest period as 12 months and the shortest period as 6 months.
CONCLUSIONSGood results can be achieved with multi-plane HA injection for rhinoplasty.
Adult ; Female ; Follow-Up Studies ; Humans ; Hyaluronic Acid ; administration & dosage ; therapeutic use ; Injections ; Male ; Rhinoplasty ; methods ; Treatment Outcome ; Young Adult
8.Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of retinoblastoma
Li-Jun, ZHOU ; Xue-Yuan, XIAO ; Kai-Li, WU ; Ju-Ling, WANG ; Hua-Sheng, YANG ; Yong-Ping, LI ; Da-Cheng, HE ; Ping, ZHANG
International Eye Science 2008;8(1):1-5
AIM:To find new biomarkers in the sera of retinoblastoma (Rb) patients with surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI TOF MS) and protein chip technique.METHODS:SELDI TOF MS, IMAC30 and CM10 protein chips were used to analyze the protein profiles from sera of 18 patients with Rb and 17 age matched controls. The protein profiling was analyzed statistically by Ciphergen protein chip software 3.0.2. The test was applied to compare the protein peak intensity. Fisher's exact test was used to compare the predominance of differential protein peaks appeared in patients.RESULTS:With IMAC30 protein chips, there were 26 proteins which appeared different in sera of patients with Rb compared to normal children. Among them, 21 proteins, I.e. 7746, 7014, 11713, 3049, 7084, 7299, 5888, 2544, 12575, 5489, 9658, 9575, 9929, 10161, 8955, 1886, 10617, 6209, 2411, 7374, 6614m/z were up regulated and 5 proteins, I.e. 8382, 7923, 7972, 8590, 66576m/z, were down regulated(P<0.01). Using the 7014 protein peak for statistical analysis, we could differentiate the patients with Rb from the healthy children with a sensitivity of 94.4% and a specificity of 82.4%. By CM10 protein chips, 4 proteins, including 3 up regulated proteins(5888, 6097, 7798 and 1 down regulated protein (8590m/z), were detected in Rb patients (P<0.01). The sensitivity and specificity were 83.3% and 70.6% respectively when 7798m/z protein peak was selected for statistical analysis.CONCLUSION:There are a few candidates as Rb biomarkers in the sera of Rb patients. SELDI TOF MS protein chip technology could be a potential method in the clinical screening test of Rb.
9.Synthesis of a supermolecular nanoparticle γ-hy-PC/Ada-Dox and its antitumor activity.
Yong-bin LI ; Kai WANG ; Tian-nan HU ; Qi-wen WANG ; Qi-da HU ; Jun ZHOU ; Xiu-rong HU ; Gu-ping TANG
Journal of Zhejiang University. Medical sciences 2012;41(6):599-609
OBJECTIVETo synthesize a (2-Hydroxypropyl)-γ-cyclodextrin-polyethylenimine/adamantane-conjugated doxorubicin (γ-hy-PC/Ada-Dox) based supramolecular nanoparticle with host-guest interaction and to identify its physicochemical characterizations and antitumor effect.
METHODSA novel non-viral gene delivery vector γ-hy-PC/Ada-Dox was synthesized based on host-guest interaction. 1H-NMR, NOESY, UV-Vis, XRD and TGA were used to confirm the structure of the vector. The DNA condensing ability of complexes was investigated by particle size, zeta potential and gel retardation assay. Cytotoxicity of complexes was determined by MTT assay in BEL-7402 and SMMC-7721 cells. Cell wound healing assay was performed in HEK293 and BEL-7404 cells. The transfection efficiency was investigated in HEK293 cells. H/E staining and cell uptake assay was performed in BEL-7402 cells.
RESULTSThe structure of γ-hy-PC/Ada-Dox was characterized by 1H-NMR, NOESY, UV-Vis, XRD, TGA. The drug loading was 0.5% and 5.5%. Gel retardation assay showed that γ-hy-PC was able to completely condense DNA at N/P ratio of 2; 0.5% and 5.5% γ-hy-PC/Ada-Dox was able to completely condense DNA at N/P ratio of 3 and 4,respectively. The cytotoxicity of polymers was lower than that of PEI25KDa. The transfection efficiency of γ-hy-PC was higher than that of γ-hy-PC/Ada-Dox at N/P ratio of 30 in HEK293 cells; and the transfection efficiency was decreasing when Ada-Dox loading was increasing. Cell uptake assay showed that γ-hy-PC/Ada-Dox was able to carry drug and FAM-siRNA into cells.
CONCLUSIONThe novel vector γ-hy-PC/Ada-Dox has been developed successfully, which has certain transfection efficiency and antitumor activity.
2-Hydroxypropyl-beta-cyclodextrin ; Adamantane ; administration & dosage ; pharmacology ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Doxorubicin ; administration & dosage ; pharmacology ; Genetic Vectors ; Humans ; Nanoparticles ; Polyethyleneimine ; Transfection ; beta-Cyclodextrins
10.Combined-modality therapy for 150 cases of early-stage Hodgkin's lymphoma.
Yi NIU ; Yuan-kai SHI ; Xiao-hui HE ; Feng-yi FENG ; Li-qiang ZHOU ; Da-zhong GU
Chinese Journal of Oncology 2008;30(8):630-634
OBJECTIVETo compare the efficacy of chemotherapy alone, radiotherapy alone and combined-modality therapy in the treatment for early-stage Hodgkin's lymphoma (HL).
METHODSFrom 1999 to 2002, totally 150 patients with stage I or II HL were treated in our hospital. They were stratified into several groups based on initial treatment strategy: chemotherapy alone (CT group, n = 22), radiotherapy alone (RT group, n = 18), combined-modality therapy (CMT group, n = 109) and surgical resection (SR group, n = 1). Chemotherapy regimens were mainly ABVD (adriamycin, bleomycin, vinblastine and dacarbazine) and MOPP (mechlorethamine, vincristine, procarbazine and prednisone). Radiotherapy modes included involved field radiotherapy (IFRT), extended field radiotherapy (EFRT) and sub-total nodal irradiation (STNI).
RESULTSThe pathological types included nodular sclerosis (NS, n = 84), mixed-cellularity (MC, n = 39), lymphocyte-predominant (LP, n = 23), lymphocyte-depleted (LD, n = 3) and nodular lymphocyte predominant Hodgkin's disease (NLPHD, n = 1). Of those, 72 were evaluble in terms of prognostic factors. No poor prognostic factor was found in 36.1% or 29.2% of the patients according to EORTC or GHSG criteria, respectively. There were 33 patients with complete response (CR), 109 with partial response (PR), 5 with stable disease (SD) and 3 with progressive disease (PD) after initial therapy. The median follow-up period was 71.5 months. The overall 7-yr survival rate was 89.3%, and treatment failure rate at 6 years was 18.8%. The response rate of CMT group was superior to that of CT group, and the patients with nodular sclerosis or mixed-cellularity type had significantly lower risk of treatment failure (P = 0.009 and 0.019, respectively). The multivariate analysis revealed that the treatment strategies affected the prognosis significantly. The risk of failure of chemotherapy alone was 2.52 times higher than that of combined-modality therapy (P = 0.004). No predictive factor affecting OS was identified by either univariate or multivariate analysis. The patients in CMT group suffered more adverse effects than those in either CT or RT groups, which mainly consisted of leucopenia, alopecia and gastrointestinal symptoms.
CONCLUSIONCombined-modality therapy is more effective than chemotherapy alone or radiotherapy alone in the treatment for early stage Hodgkin's lymphoma. Though its acute adverse effects are more severe than that of chemotherapy or radiotherapy alone, it may reduce the risk of treatment failure.
Adolescent ; Adult ; Aged ; Alopecia ; chemically induced ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Bleomycin ; adverse effects ; therapeutic use ; Child ; Child, Preschool ; Combined Modality Therapy ; Dacarbazine ; adverse effects ; therapeutic use ; Doxorubicin ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Hodgkin Disease ; drug therapy ; pathology ; radiotherapy ; Humans ; Leukopenia ; chemically induced ; Male ; Mechlorethamine ; adverse effects ; therapeutic use ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prednisone ; adverse effects ; therapeutic use ; Procarbazine ; adverse effects ; therapeutic use ; Proportional Hazards Models ; Radiotherapy ; adverse effects ; methods ; Remission Induction ; Retrospective Studies ; Survival Rate ; Vinblastine ; adverse effects ; therapeutic use ; Vincristine ; adverse effects ; therapeutic use ; Young Adult